Improvement of Cystic Fibrosis Airway Mucus  Transportability by Recombinant Human DNase Is Related to Changes in Phospholipid Profile

Zahm, Jean‐Marie; Galabert, C.; Chaffin, Alexandra; Chazalette, J. P.; Grosskopf, C.; Puchelle, Édith

doi:10.1164/ajrccm.157.6.9706036

Cited by 26 publications

(22 citation statements)

References 23 publications

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“…C, the steady state viscosity of CF sputum at shear rates 10 Ϫ2 Ϫ10 2 rad/s. Physiological rates in the normal lung are 0.2-0.9 rad/s (24). Inset, viscosities of individual CF sputum samples at physiological shear rates.…”

Section: Resultsmentioning

confidence: 99%

Enhanced Viscoelasticity of Human Cystic Fibrotic Sputum Correlates with Increasing Microheterogeneity in Particle Transport

Dawson¹,

Wirtz

Hanes³

2003

Journal of Biological Chemistry

282

291

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Current biochemical characterizations of cystic fibrosis (CF) sputum do not address the high degree of microheterogeneity in the rheological properties of the mucosal matrix and only provide bulk-average particle diffusion coefficients. The viscoelasticity of CF sputum greatly reduces the diffusion rates of colloidal particles, limiting the effectiveness of gene delivery to underlying lung cells. We determine diffusion coefficients of hundreds of individual amine-modified and carboxylated polystyrene particles (diameter 100 -500 nm) embedded in human CF sputum with 5 nm and 33 ms of spatiotemporal resolution. High resolution multiple particle tracking is used to calculate the effective viscoelastic properties of CF sputum at the micron scale, which we relate to its macroscopic viscoelasticity. CF sputum microviscosity, as probed by 100-and 200-nm particles, is an order of magnitude lower than its macroviscosity, suggesting that nanoparticles dispersed in CF sputum are transported primarily through lower viscosity pores within a highly elastic matrix. Multiple particle tracking provides a non-destructive, highly sensitive method to quantify the high heterogeneity of the mucus pore network. The mean diffusion coefficient becomes dominated by relatively few but fast-moving particles as particle size is reduced from 500 to 100 nm. Neutrally charged particles with a diameter <200 nm undergo more rapid transport in CF sputum than charged particles. Treatment with recombinant human DNase (Pulmozyme®) reduces macroviscoelastic properties of CF sputum by up to 50% and dramatically narrows the distribution of individual particle diffusion rates but surprisingly does not significantly alter the ensemble-average particle diffusion rate.

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Section: Resultsmentioning

confidence: 99%

Enhanced Viscoelasticity of Human Cystic Fibrotic Sputum Correlates with Increasing Microheterogeneity in Particle Transport

Dawson¹,

Wirtz

Hanes³

2003

Journal of Biological Chemistry

282

291

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“…Equally relevant, but a conceptually novel finding is the downregulation of DNase I, with a potential loss of capacity to break down luminal DNA, which could again increase mucus viscosity. It is notable that DNase I is used therapeutically to reduce mucus viscosity in cystic fibrosis cases by reducing the DNA content (70).…”

Section: Discussionmentioning

confidence: 99%

Expression Profiling Reveals Novel Innate and Inflammatory Responses in the Jejunal Epithelial Compartment during Infection withTrichinella spiralis

et al. 2004

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Infection with intestinal nematodes induces profound pathological changes to the gut that are associated with eventual parasite expulsion. We have applied expression profiling as an initial screening process with oligonucleotide microarrays (Affymetrix MG-U74AV2 gene chips) and time course kinetics to investigate gene transcription triggered by the intraepithelial nematode Trichinella spiralis in jejunal epithelium from BALB/c mice. Of the 4,114 genes detected, 2,617 were present in all uninfected and T. spiralis-infected replicates, 8% of which were notably upregulated, whereas 12% were downregulated at the time of worm expulsion (day 14 postinfection). Upregulation of goblet cell mucin gene transcripts intestinal mucin gene 3 (MUC3), calcium chloride channel 5 (CLCA5), and goblet cell gene 4 (GOB4) is consistent with enhanced production and alteration of mucus, whereas a 60-to 70-fold upregulation of transcripts for mast cell proteases 1 and 2 (MCPT-1 and -2) is consistent with intraepithelial mucosal mast cell recruitment. Intestinal infection can induce substantial pathological changes in the epithelial compartment (1, 37, 59). These may be preceded by specific immune responses in the lamina propria, Peyer's patches and mesenteric lymph nodes (66) and may trigger the activation of intraepithelial lymphocytes and the recruitment of inflammatory cells into the epithelial compartment (7, 39). The epithelium can itself respond to pathogens, for example, by the secretion of chloride ions and water by enterocytes (11, 51), by increased polyimmunoglobulin receptor-mediated transport of pathogen-specific immunoglobulin A (IgA) across the enterocytes (10), and by the release of mucus from goblet cells and of antimicrobial peptides from Paneth cells (37, 49). Thus, the resulting elimination of the pathogen and the associated pathology will be a complex product of both innate and pathogen-specific effector mechanisms.The intestinal nematode Trichinella spiralis dwells within epithelium and actively invades epithelial cells (36,69). This is associated with major pathological changes within the epithelial compartment such as villus atrophy and crypt hyperplasia, increased epithelial permeability, goblet cell and Paneth cell hyperplasia, and infiltration by mucosal mast cells (20,25). Some of these profound changes are immunologically mediated and are temporally associated with the rejection of adult worms (25, 33). Infection of BALB/c mice with T. spiralis is a particularly well-characterized model of infection, where parasite expulsion is partially dependent on the recruitment and activity of mucosal mast cells (28,65) Although the molecular pathology of intestinal infection is generally studied in the context of specific mechanisms, a global approach such as that afforded by transcriptomics may highlight some of the molecular pathways involved or indeed bring novel mechanisms to light. The aim of the present study was to examine, by using oligonucleotide microarrays (Affymetrix MG-U74AV2), the effect of T. spiralis on ...

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“…DNA is more widely distributed throughout purulent CF sputum samples than Ps. aeruginosa-derived alginate (Mrsny et al 1996); both recombinant human DNase (Zahm et al 1998) and alginate lyase (Mrsny et al 1994) reduce the viscosity and improve the surface properties of CF sputum, leading to speculation that aerosol delivery of such enzymes prior to chest percussion and postural drainage in severely affected CF patients could aid clearance of adhesive mucus from the airways. An alginate lyase from Bacillus circulans degraded the mucoid exopolysaccharide from the surface of CF isolates of Ps.…”

Section: Introductionmentioning

confidence: 99%

Bacteriophage-derived enzyme that depolymerizes the alginic acid capsule associated with cystic fibrosis isolates ofPseudomonas aeruginosa

Glonti

Chanishvili

Taylor³

2010

Journal of Applied Microbiology

106

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Aims: To identify enzymes associated with bacteriophages infecting cystic fibrosis (CF) strains of Pseudomonas aeruginosa that are able to degrade extracellular alginic acids elaborated by the host bacterium. Methods and Results: Plaques produced by 21 Ps. aeruginosa‐specific phages were screened for the presence of haloes, an indicator of capsule hydrolytic activity. Four phages produced haloed plaques, and one (PT‐6) was investigated further. PT‐6 was shown by electron microscopy to belong to Podoviridae family C1, to reduce the viscosity of four alginate preparations using a rolling ball viscometer and to release uronic acid‐containing fragments from the polymers, as judged by spectrophotometry and thin layer chromatography. The alginase was partially purified by gel filtration chromatography and shown to be a 37 kDa polypeptide. Conclusions: Infection of CF strains of Ps. aeruginosa by phage PT‐6 involves hydrolysis of the exopolysaccharide secreted by the host. Significance and Impact of the Study: The alginase produced by PT‐6 has the potential to increase the well‐being of CF suffers by improving the surface properties of sputum, accelerating phagocytic uptake of bacteria and perturbing bacterial growth in biofilms.

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Improvement of Cystic Fibrosis Airway Mucus Transportability by Recombinant Human DNase Is Related to Changes in Phospholipid Profile

Cited by 26 publications

References 23 publications

Enhanced Viscoelasticity of Human Cystic Fibrotic Sputum Correlates with Increasing Microheterogeneity in Particle Transport

Enhanced Viscoelasticity of Human Cystic Fibrotic Sputum Correlates with Increasing Microheterogeneity in Particle Transport

Expression Profiling Reveals Novel Innate and Inflammatory Responses in the Jejunal Epithelial Compartment during Infection withTrichinella spiralis

Bacteriophage-derived enzyme that depolymerizes the alginic acid capsule associated with cystic fibrosis isolates ofPseudomonas aeruginosa

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